Broccoli hybrid rx 05991199

ABSTRACT

The invention provides seed and plants of broccoli hybrid RX 05991199 and the parent lines thereof. The invention thus relates to the plants, seeds and tissue cultures of broccoli hybrid RX 05991199 and the parent lines thereof, and to methods for producing a broccoli plant produced by crossing such plants with themselves or with another broccoli plant, such as a plant of another genotype. The invention further relates to seeds and plants produced by such crossing. The invention further relates to parts of such plants.

FIELD OF THE INVENTION

The present invention relates to the field of plant breeding and, morespecifically, to the development of broccoli hybrid RX 05991199 and theparent broccoli lines BRM 53-3934 SI and BRM 56-3907 CMS.

BACKGROUND OF THE INVENTION

The goal of vegetable breeding is to combine various desirable traits ina single variety/hybrid. Such desirable traits may include any traitdeemed beneficial by a grower and/or consumer, including greater yield,resistance to insects or disease, tolerance to environmental stress, andnutritional value.

Breeding techniques take advantage of a plant's method of pollination.There are two general methods of pollination: a plant self-pollinates ifpollen from one flower is transferred to the same or another flower ofthe same plant or plant variety. A plant cross-pollinates if pollencomes to it from a flower of a different plant variety.

Plants that have been self-pollinated and selected for type over manygenerations become homozygous at almost all gene loci and produce auniform population of true breeding progeny, a homozygous plant. A crossbetween two such homozygous plants of different genotypes produces auniform population of hybrid plants that are heterozygous for many geneloci. Conversely, a cross of two plants each heterozygous at a number ofloci produces a population of hybrid plants that differ genetically andare not uniform. The resulting non-uniformity makes performanceunpredictable.

The development of uniform varieties requires the development ofhomozygous inbred plants, the crossing of these inbred plants, and theevaluation of the crosses. Pedigree breeding and recurrent selection areexamples of breeding methods that have been used to develop inbredplants from breeding populations. Those breeding methods combine thegenetic backgrounds from two or more plants or various other broad-basedsources into breeding pools from which new lines and hybrids derivedtherefrom are developed by selfing and selection of desired phenotypes.The new lines and hybrids are evaluated to determine which of those havecommercial potential.

SUMMARY OF THE INVENTION

In one aspect, the present invention provides a broccoli plant of thehybrid designated RX 05991199 the broccoli line BRM 53-3934 SI orbroccoli line BRM 56-3907 CMS. Also provided are broccoli plants havingall the physiological and morphological characteristics of such a plant.Parts of these broccoli plants are also provided, for example, includingpollen, an ovule, a floret, a head, and a cell of the plant.

In another aspect of the invention, a plant of broccoli hybrid RX05991199 and/or broccoli lines BRM 53-3934 SI and BRM 56-3907 CMScomprising an added heritable trait is provided. The heritable trait maycomprise a genetic locus that is, for example, a dominant or recessiveallele. In one embodiment of the invention, a plant of broccoli hybridRX 05991199 and/or broccoli lines BRM 53-3934 SI and BRM 56-3907 CMS isdefined as comprising a single locus conversion. In specific embodimentsof the invention, an added genetic locus confers one or more traits suchas, for example, herbicide tolerance, insect resistance, diseaseresistance, and modified carbohydrate metabolism. In furtherembodiments, the trait may be conferred by a naturally occurring geneintroduced into the genome of a line by backcrossing, a natural orinduced mutation, or a transgene introduced through genetictransformation techniques into the plant or a progenitor of any previousgeneration thereof. When introduced through transformation, a geneticlocus may comprise one or more genes integrated at a single chromosomallocation.

The invention also concerns the seed of broccoli hybrid RX 05991199and/or broccoli lines BRM 53-3934 SI and BRM 56-3907 CMS. The broccoliseed of the invention may be provided, in particular embodiments, as anessentially homogeneous population of broccoli seed of broccoli hybridRX 05991199 and/or broccoli lines BRM 53-3934 SI and BRM 56-3907 CMS.Essentially homogeneous populations of seed are generally free fromsubstantial numbers of other seed. Therefore, seed of hybrid RX 05991199and/or broccoli lines BRM 53-3934 SI and BRM 56-3907 CMS may beprovided, in certain embodiments of the invention, as forming at leastabout 97% of the total seed, including at least about 98%, 99% or moreof the seed. The seed population may be separately grown to provide anessentially homogeneous population of broccoli plants designated RX05991199 and/or broccoli lines BRM 53-3934 SI and BRM 56-3907 CMS.

In yet another aspect of the invention, a tissue culture of regenerablecells of a broccoli plant of hybrid RX 05991199 and/or broccoli linesBRM 53-3934 SI and BRM 56-3907 CMS is provided. The tissue culture willpreferably be capable of regenerating broccoli plants capable ofexpressing all of the physiological and morphological characteristics ofthe starting plant, and of regenerating plants having substantially thesame genotype as the starting plant. Examples of some of thephysiological and morphological characteristics of the hybrid RX05991199 and/or broccoli lines BRM 53-3934 SI and BRM 56-3907 CMSinclude those traits set forth in the tables herein. The regenerablecells in such tissue cultures may be derived, for example, from embryos,meristems, cotyledons, pollen, leaves, anthers, roots, root tips,pistils, flowers, seed and stalks. Still further, the present inventionprovides broccoli plants regenerated from a tissue culture of theinvention, the plants having all the physiological and morphologicalcharacteristics of hybrid RX 05991199 and/or broccoli lines BRM 53-3934SI and BRM 56-3907 CMS.

In still yet another aspect of the invention, processes are provided forproducing broccoli seeds, plants and parts thereof, which processesgenerally comprise crossing a first parent broccoli plant with a secondparent broccoli plant, wherein at least one of the first or secondparent broccoli plants is a plant of broccoli line BRM 53-3934 SI orbroccoli line BRM 56-3907 CMS. These processes may be furtherexemplified as processes for preparing hybrid broccoli seed or plants,wherein a first broccoli plant is crossed with a second broccoli plantof a different, distinct genotype to provide a hybrid that has, as oneof its parents, a plant of broccoli line BRM 53-3934 SI or broccoli lineBRM 56-3907 CMS. In these processes, crossing will result in theproduction of seed. The seed production occurs regardless of whether theseed is collected or not.

In one embodiment of the invention, the first step in “crossing”comprises planting seeds of a first and second parent broccoli plant,often in proximity so that pollination will occur for example, mediatedby insect vectors. Alternatively, pollen can be transferred manually.Where the plant is self-pollinated, pollination may occur without theneed for direct human intervention other than plant cultivation.

A second step may comprise cultivating or growing the seeds of first andsecond parent broccoli plants into plants that bear flowers. A thirdstep may comprise preventing self-pollination of the plants, such as byemasculating the flowers (i.e., killing or removing the pollen).

A fourth step for a hybrid cross may comprise cross-pollination betweenthe first and second parent broccoli plants. Yet another step comprisesharvesting the seeds from at least one of the parent broccoli plants.The harvested seed can be grown to produce a broccoli plant or hybridbroccoli plant.

The present invention also provides the broccoli seeds and plantsproduced by a process that comprises crossing a first parent broccoliplant with a second parent broccoli plant, wherein at least one of thefirst or second parent broccoli plants is a plant of broccoli hybrid RX05991199 and/or broccoli lines BRM 53-3934 SI and BRM 56-3907 CMS. Inone embodiment of the invention, broccoli seed and plants produced bythe process are first generation (F₁) hybrid broccoli seed and plantsproduced by crossing a plant in accordance with the invention withanother, distinct plant. The present invention further contemplatesplant parts of such an F₁ hybrid broccoli plant, and methods of usethereof. Therefore, certain exemplary embodiments of the inventionprovide an F₁ hybrid broccoli plant and seed thereof.

In still yet another aspect, the present invention provides a method ofproducing a plant derived from hybrid RX 05991199 and/or broccoli linesBRM 53-3934 SI and BRM 56-3907 CMS, the method comprising the steps of:(a) preparing a progeny plant derived from hybrid RX 05991199 and/orbroccoli lines BRM 53-3934 SI and BRM 56-3907 CMS, wherein saidpreparing comprises crossing a plant of the hybrid RX 05991199 and/orbroccoli lines BRM 53-3934 SI and BRM 56-3907 CMS with a second plant;and (b) crossing the progeny plant with itself or a second plant toproduce a seed of a progeny plant of a subsequent generation. In furtherembodiments, the method may additionally comprise: (c) growing a progenyplant of a subsequent generation from said seed of a progeny plant of asubsequent generation and crossing the progeny plant of a subsequentgeneration with itself or a second plant; and repeating the steps for anadditional 3-10 generations to produce a plant derived from hybrid RX05991199 and/or broccoli lines BRM 53-3934 SI and BRM 56-3907 CMS. Theplant derived from hybrid RX 05991199 and/or broccoli lines BRM 53-3934SI and BRM 56-3907 CMS may be an inbred line, and the aforementionedrepeated crossing steps may be defined as comprising sufficientinbreeding to produce the inbred line. In the method, it may bedesirable to select particular plants resulting from step (c) forcontinued crossing according to steps (b) and (c). By selecting plantshaving one or more desirable traits, a plant derived from hybrid RX05991199 and/or broccoli lines BRM 53-3934 SI and BRM 56-3907 CMS isobtained which possesses some of the desirable traits of the line/hybridas well as potentially other selected traits.

In certain embodiments, the present invention provides a method ofproducing food or feed comprising: (a) obtaining a plant of broccolihybrid RX 05991199 and/or broccoli lines BRM 53-3934 SI and BRM 56-3907CMS, wherein the plant has been cultivated to maturity, and (b)collecting tissue of the plant.

In still yet another aspect of the invention, the genetic complement ofbroccoli hybrid RX 05991199 and/or broccoli lines BRM 53-3934 SI and BRM56-3907 CMS is provided. The phrase “genetic complement” is used torefer to the aggregate of nucleotide sequences, the expression of whichsequences defines the phenotype of, in the present case, a broccoliplant, or a cell or tissue of that plant. A genetic complement thusrepresents the genetic makeup of a cell, tissue or plant, and a hybridgenetic complement represents the genetic make up of a hybrid cell,tissue or plant. The invention thus provides broccoli plant cells thathave a genetic complement in accordance with the broccoli plant cellsdisclosed herein, and plants, seeds and plants containing such cells.

Plant genetic complements may be assessed by genetic marker profiles,and by the expression of phenotypic traits that are characteristic ofthe expression of the genetic complement, e.g., isozyme typing profiles.It is understood that hybrid RX 05991199 and/or broccoli lines BRM53-3934 SI and BRM 56-3907 CMS could be identified by any of the manywell known techniques such as, for example, Simple Sequence LengthPolymorphisms (SSLPs) (Williams et al., 1990), Randomly AmplifiedPolymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF),Sequence Characterized Amplified Regions (SCARs), Arbitrary PrimedPolymerase Chain Reaction (AP-PCR), Amplified Fragment LengthPolymorphisms (AFLPs) (EP 534 858, specifically incorporated herein byreference in its entirety), and Single Nucleotide Polymorphisms (SNPs)(Wang et al., 1998).

In still yet another aspect, the present invention provides hybridgenetic complements, as represented by broccoli plant cells, tissues,plants, and seeds, formed by the combination of a haploid geneticcomplement of a broccoli plant of the invention with a haploid geneticcomplement of a second broccoli plant, preferably, another, distinctbroccoli plant. In another aspect, the present invention provides abroccoli plant regenerated from a tissue culture that comprises a hybridgenetic complement of this invention.

In still yet another aspect, the invention provides a method ofdetermining the genotype of a plant of broccoli hybrid RX 05991199and/or broccoli lines BRM 53-3934 SI and BRM 56-3907 CMS comprisingdetecting in the genome of the plant at least a first polymorphism. Themethod may, in certain embodiments, comprise detecting a plurality ofpolymorphisms in the genome of the plant. The method may furthercomprise storing the results of the step of detecting the plurality ofpolymorphisms on a computer readable medium. The invention furtherprovides a computer readable medium produced by such a method.

Any embodiment discussed herein with respect to one aspect of theinvention applies to other aspects of the invention as well, unlessspecifically noted.

The term “about” is used to indicate that a value includes the standarddeviation of the mean for the device or method being employed todetermine the value. The use of the term “or” in the claims is used tomean “and/or” unless explicitly indicated to refer to alternatives onlyor the alternatives are mutually exclusive. When used in conjunctionwith the word “comprising” or other open language in the claims, thewords “a” and “an” denote “one or more,” unless specifically notedotherwise. The terms “comprise,” “have” and “include” are open-endedlinking verbs. Any forms or tenses of one or more of these verbs, suchas “comprises,” “comprising,” “has,” “having,” “includes” and“including,” are also open-ended. For example, any method that“comprises,” “has” or “includes” one or more steps is not limited topossessing only those one or more steps and also covers other unlistedsteps. Similarly, any plant that “comprises,” “has” or “includes” one ormore traits is not limited to possessing only those one or more traitsand covers other unlisted traits.

Other objects, features and advantages of the present invention willbecome apparent from the following detailed description. It should beunderstood, however, that the detailed description and any specificexamples provided, while indicating specific embodiments of theinvention, are given by way of illustration only, since various changesand modifications within the spirit and scope of the invention willbecome apparent to those skilled in the art from this detaileddescription.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides methods and compositions relating to plants,seeds and derivatives of RX 05991199 broccoli hybrid RX 05991199 and/orbroccoli lines BRM 53-3934 SI and BRM 56-3907 CMS. The hybrid RX05991199 was produced by the cross of parent lines BRM 53-3934 SI andBRM 56-3907 CMS. The parent lines show uniformity and stability withinthe limits of environmental influence. By crossing the parent lines,uniform plants of hybrid RX 05991199 can be obtained.

The parents of hybrid RX 05991199 are BRM 53-3934 SI and BRM 56-3907CMS. The parent BRM 56-3907 CMS is a known inbred that served as theparent of the commercial hybrid “Ironman”. BRM 56-3907 CMS is thesubject of, and is described in, EU Plant Variety Rights Certificate#20341, granted Jun. 18, 2007.

In accordance with one aspect of the present invention, there isprovided a plant having the physiological and morphologicalcharacteristics of broccoli hybrid RX 05991199 and the parent linesthereof. A description of the physiological and morphologicalcharacteristics of such plants is presented in Tables 1-2.

TABLE 1 Physiological and Morphological Characteristics of Hybrid RX05991199 Characteristic RX 05991199CMS Ironman A Region of Adaptation NWEurope NW Europe B Maturity, Spring Planted days from direct seeding tono direct seeding no direct seeding 50% harvest days from transplantingto 50% 64 63 harvest transplant date 10-May-10 10-May-10 length ofharvest period in days  8  6 first harvest date 12-Jul-10 12-Jul-10 lastharvest date 19-Jul-10 17-Jul-10 harvest season (main crop at summersummer 50% harvest) time of harvest maturity (50% medium (Sumosun)medium (Sumosun) of plants) time of beginning of flowering medium*8(Coaster, medium*8 (Coaster, (50% of plants with at least Cruiser)Cruiser) 10% flowers) **choice for UPOV TG only C Seedling cotyledoncolor medium green medium green RHS color chart value for 147B 147Bseedling cotyledon color cotyledon anthocyanin intermediate intermediatehypocotyl anthocyanin strong strong D Plant plant height in centimeters56.3 cm 61.0 cm from soil line to top of leaves head height incentimeters from 33.7 cm 34.7 cm soil line to top of leaves height atharvest maturity medium (Coaster) medium (Coaster) number of stems one(Ramoso Calabrese, one (Ramoso Shogun) Calabrese, Shogun) branchesmedium few habit intermediate intermediate market class fresh marketfresh market life cycle annual annual type of variety first generationhybrid first generation hybrid E Leaves outer leaves: number of leaves14 17 per plant (at harvest) outer leaves: width (at 22.6 cm 24.2 cmmidpoint of plant including petiole) leaf: width medium (Buccaneer,medium (Buccaneer, Green Belt) Green Belt) outer leaves: length (at 48.7cm 53.2 cm midpoint of plant including petiole) leaf: length (includingpetiole) medium (Brigadeer, long (Green Duke, Sumosun) Laser) outerleaves: petiole length 22.1 cm 24.9 cm petiole: length medium (Emperor,long (Groene Ramoso Calabrese) Calabrese, Premium Crop) outer leaves:leaf ratio- 2:1 2:1 length/width outer leaves: leaf attachment petiolatepetiolate outer leaves: wax presence weak intermediate leaf: number oflobes medium (Coaster, medium (Coaster, Topper) Topper) outer leaves:foliage color grey green grey green (with wax, if present) outer leaves:foliage color 189A 189A (with wax, if present; RHS color chart value)leaf blade: color grey green (Bishop) grey green (Bishop) leaf blade:intensity of color dark dark leaf blade: anthocyanin absent (Claudia,absent (Claudia, coloration Embassy) Embassy) leaf blade: undulation ofweak (Beaufort, Early weak (Beaufort, margin Pack, Laser, Paladin) EarlyPack, Laser, Paladin) leaf blade: dentation of margin weak (Galaxy) weak(Galaxy) outer leaves: leaf shape elliptic elliptic outer leaves: leafbase blunt blunt outer leaves: leaf apex blunt blunt outer leaves: leafmargins slightly wavy slightly wavy outer leaves: leaf veinsintermediate intermediate outer leaves: midrib slightly raised notraised leaf blade: blistering medium (Medium Late medium (Medium 145,Skiff) Late 145, Skiff) outer leaves: attitude (leaf semi-erect (35-55semi-erect (35-55 angle from ground) degrees) degrees) leaf: attitude(at beginning of semi-erect (Arcadia, semi-erect head formation) Asti,Civet, Claudia) outer leaves: torsion of leaf tip none none outerleaves: profile of upper planar planar side of leaf F Head length ofbranching at base very short (Viola) very short (Viola) (excluding stem)diameter at widest point (at 15.2 cm 14.7 cm market maturity) depth (atmarket maturity) 9.7 cm 9.4 cm weight, market trimmed (at 439.9 gm 400.3gm market maturity) color grey green (Brigadeer, grey green Galaxy)(Brigadeer, Galaxy) intensity of color medium medium RHS color chartvalue for head 189B 189B color anthocyanin coloration absent (EarlyWhite present (Brigadeer, Sprouting) Shogun, Viola) shape (at marketmaturity) transverse elliptic transverse elliptic (Buccaneer, Futura)(Buccaneer, Futura) dome shape (at market domed domed maturity) size (atmarket maturity) for medium** (Dundee, medium** (Dundee, US Exhibit Conly **choice Early Man) Early Man) compactness/firmness (at medium(Late Corona) short pedicels/tight/ market maturity) firm (Captain)surface knobbling (at market fine (Apollo, Brigadeer) medium (Southernmaturity) Comet) texture medium (Clipper, fine (Auriga, Bishop, Coaster)Green Top) bead size (at market maturity) medium small flower buds (atmarket even in size even in size maturity) anthocyanin coloration ofleaf absent absent axils (at market maturity) anthocyanin coloration ofleaf absent absent veins (at market maturity) anthocyanin coloration ofleaf absent absent blade (at market maturity) anthocyanin coloration ofabsent absent entire plant (at market maturity) anthocyanin colorationof leaf absent (Claudia, absent (Claudia, petiole (at market maturity)Embassy) Embassy) color of head leaves (at market green green maturity)RHS color chart value for the N189B N189B color of head leaves bractsabsent (Gem, Orion) absent (Gem, Orion) secondary heads (at marketcombination, present** present, combination maturity) (Marathon,Tribute, Late Purple Sprouting) prominence of secondary headsintermediate (Citation) weak (at market maturity) number of secondaryheads (at  4  2 market maturity) G Flower Color **choice for UPOV TGyellow** (Brigadeer, yellow** (Brigadeer, only Orion) Orion) intensityof yellow color medium (Capitol, medium (Capitol, Corvet) Corvet) color4B 4B stalk color green green RHS color chart value for 138B 138B flowerstalk color male sterility present (Chevalier, present (Chevalier,Montop) Montop) *These are typical values. Values may vary due toenvironment. Other values that are substantially equivalent are alsowithin the scope of the invention.

TABLE 2 Physiological and Morphological Characteristics of Line BRM53-3934 SI Characteristic BMR53-3934SI Sibsey A Region of Adaptation NWEurope NW Europe B Maturity, Spring Planted days from direct seeding tono direct seeding no direct seeding 50% harvest days from transplantingto 50% 76 54 harvest transplant date 10-May-10 10-May-10 length ofharvest period in days 16 5 first harvest date 17-Jul-10 1-Jul-10 lastharvest date 1-Aug-10 5-Jul-10 harvest season (main crop at summersummer 50% harvest) time of harvest maturity (50% very late (Late Purplevery early of plants) Sprouting) (Earlyman, Primor) time of beginning offlowering late** (Shogun, Viola) early** (Clipper, (50% of plants withat least Southern Comet) 10% flowers) **choice for UPOV TG only CSeedling cotyledon color medium green medium green RHS color chart valuefor 147B 147B seedling cotyledon color cotyledon anthocyanin weakintermediate hypocotyl anthocyanin weak intermediate D Plant plantheight in centimeters 51.7 cm 55.0 cm from soil line to top of leaveshead height in centimeters from 30.3 cm 50.0 cm soil line to top ofleaves height at harvest maturity medium (Coaster) medium (Coaster)number of stems one (Ramoso Calabrese, one (Ramoso Shogun) Calabrese,Shogun) branches medium many habit compact compact market class freshmarket fresh market life cycle annual annual type of variety inbredfirst generation hybrid E Leaves outer leaves: number of leaves 20 12per plant (at harvest) outer leaves: width (at 21.8 cm 18.9 cm midpointof plant including petiole) leaf: width medium (Buccaneer, narrow(Arcadia, Green Belt) Brigadeer) outer leaves: length (at 45.3 cm 39.1cm midpoint of plant including petiole) leaf: length (including petiole)medium (Brigadeer, short (Dandy Early, Sumosun) Emperor) outer leaves:petiole length 20.2 cm 16.0 cm petiole: length medium (Emperor, short(High Sierra, Ramoso Calabrese) Padovano) outer leaves: leaf ratio- 2:12:1 length/width outer leaves: leaf attachment petiolate petiolate outerleaves: wax presence intermediate intermediate leaf: number of lobesmedium (Coaster, medium (Coaster, Topper) Topper) outer leaves: foliagecolor blue green grey green (with wax, if present) outer leaves: foliagecolor N198A 189A (with wax, if present; RHS color chart value) leafblade: color blue green (Citation, grey green (Bishop) Esquire,Symphony) leaf blade: intensity of color medium medium leaf blade:anthocyanin absent (Claudia, absent (Claudia, coloration Embassy)Embassy) leaf blade: undulation of weak (Beaufort, Early strong (Aikido,margin Pack, Laser, Paladin) Marathon, Samurai) leaf blade: dentation ofmargin weak (Galaxy) weak (Galaxy) outer leaves: leaf shape ellipticelliptic outer leaves: leaf base blunt blunt outer leaves: leaf apexblunt blunt outer leaves: leaf margins slightly wavy very wavy outerleaves: leaf veins intermediate thin outer leaves: midrib slightlyraised not raised leaf blade: blistering absent or very weak absent orvery weak (Buccaneer, Colibri) (Buccaneer, Colibri) outer leaves:attitude (leaf semi-erect (35-55 semi-erect (35-55 angle from ground)degrees) degrees) leaf: attitude (at beginning of semi-erect (Arcadia,semi-erect head formation) Asti, Civet, Claudia) outer leaves: torsionof leaf tip none weak outer leaves: profile of upper concave convex sideof leaf F Head length of branching at base short (Brigadeer, medium(Capitol, (excluding stem) Buccaneer Emperor) Green Duke, Perseus)diameter at widest point (at 11.4 cm 13.7 cm market maturity) depth (atmarket maturity) 10.4 cm 10.9 cm weight, market trimmed (at 164.3 gm182.7 gm market maturity) color light green; grey-green grey-green(Brigadeer, Galaxy) (Brigadeer, Galaxy) intensity of color medium mediumRHS color chart value for head 189A/144A N189B color anthocyanincoloration absent (Early White absent (Early White Sprouting) Sprouting)shape (at market maturity) circular (Esquire) transverse elliptic(Buccaneer, Futura) dome shape (at market domed domed maturity) size (atmarket maturity) for US very small (Early Purple small** (Orbit, ExhibitC only **choice Sprouting) Scorpio) compactness/firmness (at longpedicels/loose medium (Late market maturity) (Caravel) Corona) surfaceknobbling (at market medium (Southern fine (Apollo, maturity) Comet)Brigadeer) texture fine (Auriga, Bishop, fine (Auriga, Bishop, GreenTop); coarse Green Top) (Citation) bead size (at market maturity)small/medium/large small flower buds (at market uneven in size (cateye)even in size maturity) anthocyanin coloration of leaf absent absentaxils (at market maturity) anthocyanin coloration of leaf absent absentveins (at market maturity) anthocyanin coloration of leaf absent absentblade (at market maturity) anthocyanin coloration of entire absentabsent plant (at market maturity) anthocyanin coloration of leaf absent(Claudia, absent (Claudia, petiole (at market maturity) Embassy)Embassy) color of head leaves (at market green green maturity) RHS colorchart value for the N189B 189A color of head leaves bracts present(Ramoso absent (Gem, Orion) Calabrese) secondary heads (at market basal,present** combination maturity) (Marathon, Tribute, Late PurpleSprouting) prominence of secondary heads intermediate (Citation)intermediate (at market maturity) (Citation) number of secondary heads(at  3  4 market maturity) G Flower Color **choice for UPOV TG yellow**(Brigadeer, yellow** (Brigadeer, only Orion) Orion) intensity of yellowcolor dark (Gem, Orion) dark (Gem, Orion) color 4A 4A stalk color greengreen RHS color chart value for 138B 138B flower stalk color malesterility absent (Marathon) present (Chevalier, Montop) *These aretypical values. Values may vary due to environment. Other values thatare substantially equivalent are also within the scope of the invention.

The MSB content of hybrid RX 05991199 relative to the hybrid “Heritage”was the subject of an objective analysis. The results of the analysisare presented below.

TABLE 3 Analysis of MSB Content of RX 05991199 Least-Squares Means forMSB Varieties (micromoles/gm/FW) RX 05991199 3.5833927 Heritage1.6311887

A head-to-head analysis was also made of glucoraphanin (MSB) content ofRX 05991199 relative to the hybrid Ironman in multiple locations andplantings. The results are presented below.

TABLE 4 Analysis of Glucoraphanin Content of RX 05991199 Relative toHybrid Ironman Sub- Plant- calculated in μmol per gDW Variety CountryRegion ing Glucoiberin Glucoraphanin Ironman Italy South A 2.1 3.9 RX05991199 Italy South A 5.0 20.0 RX 05991199 Italy South A 1.5 6.7Ironman Spain South B 2.1 3.1 RX 05991199 Spain South B 3.5 13.4 IronmanItaly South C 3.2 9.2 Ironman Italy South C 3.6 9.1 Ironman Italy SouthC 3.4 10.3 Ironman Italy South C 3.3 10.8 RX 05991199 Italy South C 4.720.7 RX 05991199 Italy South C 5.4 22.3 RX 05991199 Italy South C 5.526.0 RX 05991199 Italy South C 4.7 22.9 Ironman Spain South D 1.9 5.5Ironman Spain South D 2.8 9.6 Ironman Spain South D 2.4 5.9 RX 05991199Spain South D 5.1 21.7 RX 05991199 Spain South D 5.5 24.8 RX 05991199Spain South D 5.1 25.7 Ironman UK North A 2.5 9.8 Ironman UK North A 3.112.8 RX 05991199 UK North A 3.8 21.3 RX 05991199 UK North A 3.4 24.4Ironman UK North B 0.9 7.1 Ironman UK North B 1.9 12.1 RX 05991199 UKNorth B 2.4 11.9 RX 05991199 UK North B 3.0 16.0 Ironman UK North C 1.85.8 Ironman UK North C 1.9 5.8 RX 05991199 UK North C 3.1 14.1 RX05991199 UK North C 3.3 15.1 Ironman UK North D 1.1 6.2 Ironman UK NorthD 0.9 6.2 RX 05991199 UK North D 3.2 17.2 RX 05991199 UK North D 3.116.6 ironman Italy South F 0.9 5.1 ironman Italy South F 1.0 5.3 RX05991199 Italy South F 3.8 24.5 RX 05991199 Italy South F 3.6 25.1ironman Italy South A 0.9 4.3 ironman Italy South A 1.0 5.1 RX 05991199Italy South A 4.6 23.5 RX 05991199 Italy South A 4.0 22.4 ironman SpainSouth B 1.5 6.0 ironman Spain South B 2.4 11.2 RX 05991199 Spain South B4.0 20.7 RX 05991199 Spain South B 4.4 22.1 ironman Spain South C 1.98.3 ironman Spain South C 1.7 8.8 RX 05991199 Spain South C 4.1 24.3 RX05991199 Spain South C 4.3 22.4 Ironman Spain South D 1.2 5.5 IronmanSpain South D 0.9 4.9 RX 05991199 Spain South D 3.5 19.0 RX 05991199Spain South D 2.8 17.5 Ironman Italy South E 0.9 7.6 Ironman Italy SouthE 0.8 6.7 RX 05991199 Italy South E 3.0 24.4 RX 05991199 Italy South E1.8 23.3 Ironman Spain South F 0.8 5.0 Ironman Spain South F 0.6 3.8 RX05991199 Spain South F 2.9 17.1 RX 05991199 Spain South F 3.0 19.3Ironman Spain South G 1.0 6.3 Ironman Spain South G 0.9 6.2 RX 05991199Spain South G 3.4 20.8 RX 05991199 Spain South G 3.0 19.9 Ironman SpainSouth H 2.5 14.7 Ironman Spain South H 2.3 14.1 RX 05991199 Spain SouthH 3.6 21.9 RX 05991199 Spain South H 4.0 23.0 Ironman UK North B 0.9 6.2Ironman UK North B 1.0 6.1 RX 05991199 UK North B 4.2 20.9 RX 05991199UK North B 3.6 18.6 Ironman UK North C 2.1 6.3 Ironman UK North C 1.96.4 RX 05991199 UK North C 4.7 15.1 RX 05991199 UK North C 4.4 15.5Ironman UK North D 1.7 11.3 Ironman UK North D 1.1 6.3 RX 05991199 UKNorth D 4.1 22.0 RX 05991199 UK North D 2.7 14.6 Ironman UK North E 2.111.9 RX 05991199 UK North E 5.3 24.2 Ironman UK North F 1.2 8.0 IronmanUK North F 1.5 8.7 RX 05991199 UK North F 4.2 19.9 RX 05991199 UK NorthF 4.5 21.9 Ironman UK North G 1.0 6.1 Ironman UK North G 1.0 6.7 RX05991199 UK North G 2.7 16.8 RX 05991199 UK North G 2.8 17.7 Ironman UKNorth H 1.0 6.7 Ironman UK North H 0.8 5.8 RX 05991199 UK North H 3.419.2 RX 05991199 UK North H 3.9 23.1 Ironman Spain South A 0.6 4.0Ironman Spain South A 0.9 4.2 Ironman Spain South A 0.8 3.7 RX 05991199Spain South A 2.6 13.3 RX 05991199 Spain South A 2.7 14.1 RX 05991199Spain South A 2.8 13.4 Ironman Italy South B 1.4 7.3 Ironman Italy SouthB 1.2 8.2 RX 05991199 Italy South B 4.9 26.6 RX 05991199 Italy South B3.5 18.7 Ironman Spain South C 1.1 5.9 Ironman Spain South C 1.0 4.8Ironman Spain South C 1.0 5.5 RX 05991199 Spain South C 3.3 16.0 RX05991199 Spain South C 2.7 13.0 RX 05991199 Spain South C 2.9 11.7Ironman Italy South D 1.1 5.9 Ironman Italy South D 1.1 5.3 RX 05991199Italy South D 2.9 16.0 RX 05991199 Italy South D 3.9 21.4 Ironman SpainSouth E 0.9 4.3 Ironman Spain South E 1.0 3.9 Ironman Spain South E 1.14.8 RX 05991199 Spain South E 3.6 21.2 RX 05991199 Spain South E 2.216.5 RX 05991199 Spain South E 2.5 15.5 Ironman Spain South F 1.1 4.6Ironman Spain South F 0.8 4.4 Ironman Spain South F 1.0 4.5 RX 05991199Spain South F 1.5 9.7 RX 05991199 Spain South F 2.3 15.4 RX 05991199Spain South F 2.4 15.3 Ironman Spain South G 1.2 4.4 Ironman Spain SouthG 1.4 5.6 Ironman Spain South G 1.0 3.4 RX 05991199 Spain South G 2.916.7 RX 05991199 Spain South G 2.8 18.7 RX 05991199 Spain South G 3.015.9 Ironman UK North L 0.7 8.2 Ironman UK North L 0.8 9.8 Ironman UKNorth L 0.8 10.3 RX 05991199 UK North L 1.6 15.0 RX 05991199 UK North L1.9 17.1 RX 05991199 UK North L 1.9 18.1 Ironman UK North M 0.2 4.9Ironman UK North M 0.3 4.5 Ironman UK North M 0.3 5.2 RX 05991199 UKNorth M 1.1 8.5 RX 05991199 UK North M 1.6 9.8 RX 05991199 UK North M1.3 10.0

A. Breeding Broccoli Plants

One aspect of the current invention concerns methods for producing seedof broccoli hybrid RX 05991199 involving crossing broccoli lines BRM53-3934 SI and BRM 56-3907 CMS. Alternatively, in other embodiments ofthe invention, hybrid RX 05991199, line BRM 53-3934 SI, or line BRM56-3907 CMS may be crossed with itself or with any second plant. Suchmethods can be used for propagation of hybrid RX 05991199 and/or thebroccoli lines BRM 53-3934 SI and BRM 56-3907 CMS, or can be used toproduce plants that are derived from hybrid RX 05991199 and/or thebroccoli lines BRM 53-3934 SI and BRM 56-3907 CMS. Plants derived fromhybrid RX 05991199 and/or the broccoli lines BRM 53-3934 SI and BRM56-3907 CMS may be used, in certain embodiments, for the development ofnew broccoli varieties.

The development of new varieties using one or more starting varieties iswell known in the art. In accordance with the invention, novel varietiesmay be created by crossing hybrid RX 05991199 followed by multiplegenerations of breeding according to such well known methods. Newvarieties may be created by crossing with any second plant. In selectingsuch a second plant to cross for the purpose of developing novel lines,it may be desired to choose those plants which either themselves exhibitone or more selected desirable characteristics or which exhibit thedesired characteristic(s) when in hybrid combination. Once initialcrosses have been made, inbreeding and selection take place to producenew varieties. For development of a uniform line, often five or moregenerations of selfing and selection are involved.

Uniform lines of new varieties may also be developed by way ofdouble-haploids. This technique allows the creation of true breedinglines without the need for multiple generations of selfing andselection. In this manner true breeding lines can be produced in aslittle as one generation. Haploid embryos may be produced frommicrospores, pollen, anther cultures, or ovary cultures. The haploidembryos may then be doubled autonomously, or by chemical treatments(e.g. colchicine treatment). Alternatively, haploid embryos may be growninto haploid plants and treated to induce chromosome doubling. In eithercase, fertile homozygous plants are obtained. In accordance with theinvention, any of such techniques may be used in connection with a plantof the invention and progeny thereof to achieve a homozygous line.

Backcrossing can also be used to improve an inbred plant. Backcrossingtransfers a specific desirable trait from one inbred or non-inbredsource to an inbred that lacks that trait. This can be accomplished, forexample, by first crossing a superior inbred (A) (recurrent parent) to adonor inbred (non-recurrent parent), which carries the appropriate locusor loci for the trait in question. The progeny of this cross are thenmated back to the superior recurrent parent (A) followed by selection inthe resultant progeny for the desired trait to be transferred from thenon-recurrent parent. After five or more backcross generations withselection for the desired trait, the progeny have the characteristicbeing transferred, but are like the superior parent for most or almostall other loci. The last backcross generation would be selfed to givepure breeding progeny for the trait being transferred.

The plants of the present invention are particularly well suited for thedevelopment of new lines based on the elite nature of the geneticbackground of the plants. In selecting a second plant to cross with RX05991199 and/or broccoli lines BRM 53-3934 SI and BRM 56-3907 CMS forthe purpose of developing novel broccoli lines, it will typically bepreferred to choose those plants which either themselves exhibit one ormore selected desirable characteristics or which exhibit the desiredcharacteristic(s) when in hybrid combination. Examples of desirabletraits may include, in specific embodiments, high seed yield, high seedgermination, seedling vigor, high yield, disease tolerance orresistance, and adaptability for soil and climate conditions.Consumer-driven traits, such as a head shape, nutritional value, andtaste are other examples of traits that may be incorporated into newlines of broccoli plants developed by this invention.

B. Further Embodiments of the Invention

In certain aspects of the invention, plants described herein areprovided modified to include at least a first desired heritable trait.Such plants may, in one embodiment, be developed by a plant breedingtechnique called backcrossing, wherein essentially all of themorphological and physiological characteristics of a variety arerecovered in addition to a genetic locus transferred into the plant viathe backcrossing technique. The term single locus converted plant asused herein refers to those broccoli plants which are developed by aplant breeding technique called backcrossing, wherein essentially all ofthe morphological and physiological characteristics of a variety arerecovered in addition to the single locus transferred into the varietyvia the backcrossing technique. By essentially all of the morphologicaland physiological characteristics, it is meant that the characteristicsof a plant are recovered that are otherwise present when compared in thesame environment, other than an occasional variant trait that mightarise during backcrossing or direct introduction of a transgene.

Backcrossing methods can be used with the present invention to improveor introduce a characteristic into the present variety. The parentalbroccoli plant which contributes the locus for the desiredcharacteristic is termed the nonrecurrent or donor parent. Thisterminology refers to the fact that the nonrecurrent parent is used onetime in the backcross protocol and therefore does not recur. Theparental broccoli plant to which the locus or loci from the nonrecurrentparent are transferred is known as the recurrent parent as it is usedfor several rounds in the backcrossing protocol.

In a typical backcross protocol, the original variety of interest(recurrent parent) is crossed to a second variety (nonrecurrent parent)that carries the single locus of interest to be transferred. Theresulting progeny from this cross are then crossed again to therecurrent parent and the process is repeated until a broccoli plant isobtained wherein essentially all of the morphological and physiologicalcharacteristics of the recurrent parent are recovered in the convertedplant, in addition to the single transferred locus from the nonrecurrentparent.

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute a single trait or characteristic in the originalvariety. To accomplish this, a single locus of the recurrent variety ismodified or substituted with the desired locus from the nonrecurrentparent, while retaining essentially all of the rest of the desiredgenetic, and therefore the desired physiological and morphologicalconstitution of the original variety. The choice of the particularnonrecurrent parent will depend on the purpose of the backcross; one ofthe major purposes is to add some commercially desirable trait to theplant. The exact backcrossing protocol will depend on the characteristicor trait being altered and the genetic distance between the recurrentand nonrecurrent parents. Although backcrossing methods are simplifiedwhen the characteristic being transferred is a dominant allele, arecessive allele, or an additive allele (between recessive anddominant), may also be transferred. In this instance it may be necessaryto introduce a test of the progeny to determine if the desiredcharacteristic has been successfully transferred.

In one embodiment, progeny broccoli plants of a backcross in which aplant described herein is the recurrent parent comprise (i) the desiredtrait from the non-recurrent parent and (ii) all of the physiologicaland morphological characteristics of the recurrent parent as determinedat the 5% significance level when grown in the same environmentalconditions.

New varieties can also be developed from more than two parents. Thetechnique, known as modified backcrossing, uses different recurrentparents during the backcrossing. Modified backcrossing may be used toreplace the original recurrent parent with a variety having certain moredesirable characteristics or multiple parents may be used to obtaindifferent desirable characteristics from each.

Many single locus traits have been identified that are not regularlyselected for in the development of a new inbred but that can be improvedby backcrossing techniques. Single locus traits may or may not betransgenic; examples of these traits include, but are not limited to,herbicide resistance, resistance to bacterial, fungal, or viral disease,insect resistance, modified fatty acid or carbohydrate metabolism, andaltered nutritional quality. These comprise genes generally inheritedthrough the nucleus.

Direct selection may be applied where the single locus acts as adominant trait. For this selection process, the progeny of the initialcross are assayed for viral resistance and/or the presence of thecorresponding gene prior to the backcrossing. Selection eliminates anyplants that do not have the desired gene and resistance trait, and onlythose plants that have the trait are used in the subsequent backcross.This process is then repeated for all additional backcross generations.

Selection of broccoli plants for breeding is not necessarily dependenton the phenotype of a plant and instead can be based on geneticinvestigations. For example, one can utilize a suitable genetic markerwhich is closely genetically linked to a trait of interest. One of thesemarkers can be used to identify the presence or absence of a trait inthe offspring of a particular cross, and can be used in selection ofprogeny for continued breeding. This technique is commonly referred toas marker assisted selection. Any other type of genetic marker or otherassay which is able to identify the relative presence or absence of atrait of interest in a plant can also be useful for breeding purposes.Procedures for marker assisted selection are well known in the art. Suchmethods will be of particular utility in the case of recessive traitsand variable phenotypes, or where conventional assays may be moreexpensive, time consuming or otherwise disadvantageous. Types of geneticmarkers which could be used in accordance with the invention include,but are not necessarily limited to, Simple Sequence Length Polymorphisms(SSLPs) (Williams et al., 1990), Randomly Amplified Polymorphic DNAs(RAPDs), DNA Amplification Fingerprinting (DAF), Sequence CharacterizedAmplified Regions (SCARs), Arbitrary Primed Polymerase Chain Reaction(AP-PCR), Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858,specifically incorporated herein by reference in its entirety), andSingle Nucleotide Polymorphisms (SNPs) (Wang et al., 1998).

C. Plants Derived by Genetic Engineering

Many useful traits that can be introduced by backcrossing, as well asdirectly into a plant, are those which are introduced by genetictransformation techniques. Genetic transformation may therefore be usedto insert a selected transgene into a plant of the invention or may,alternatively, be used for the preparation of transgenes which can beintroduced by backcrossing. Methods for the transformation of plantsthat are well known to those of skill in the art and applicable to manycrop species include, but are not limited to, electroporation,microprojectile bombardment, Agrobacterium-mediated transformation anddirect DNA uptake by protoplasts.

To effect transformation by electroporation, one may employ eitherfriable tissues, such as a suspension culture of cells or embryogeniccallus or alternatively one may transform immature embryos or otherorganized tissue directly. In this technique, one would partiallydegrade the cell walls of the chosen cells by exposing them topectin-degrading enzymes (pectolyases) or mechanically wound tissues ina controlled manner.

An efficient method for delivering transforming DNA segments to plantcells is microprojectile bombardment. In this method, particles arecoated with nucleic acids and delivered into cells by a propellingforce. Exemplary particles include those comprised of tungsten,platinum, and preferably, gold. For the bombardment, cells in suspensionare concentrated on filters or solid culture medium. Alternatively,immature embryos or other target cells may be arranged on solid culturemedium. The cells to be bombarded are positioned at an appropriatedistance below the macroprojectile stopping plate.

An illustrative embodiment of a method for delivering DNA into plantcells by acceleration is the Biolistics Particle Delivery System, whichcan be used to propel particles coated with DNA or cells through ascreen, such as a stainless steel or Nytex screen, onto a surfacecovered with target cells. The screen disperses the particles so thatthey are not delivered to the recipient cells in large aggregates.Microprojectile bombardment techniques are widely applicable, and may beused to transform virtually any plant species.

Agrobacterium-mediated transfer is another widely applicable system forintroducing gene loci into plant cells. An advantage of the technique isthat DNA can be introduced into whole plant tissues, thereby bypassingthe need for regeneration of an intact plant from a protoplast. ModernAgrobacterium transformation vectors are capable of replication in E.coli as well as Agrobacterium, allowing for convenient manipulations(Klee et al., 1985). Moreover, recent technological advances in vectorsfor Agrobacterium-mediated gene transfer have improved the arrangementof genes and restriction sites in the vectors to facilitate theconstruction of vectors capable of expressing various polypeptide codinggenes. The vectors described have convenient multi-linker regionsflanked by a promoter and a polyadenylation site for direct expressionof inserted polypeptide coding genes. Additionally, Agrobacteriumcontaining both armed and disarmed Ti genes can be used fortransformation.

In those plant strains where Agrobacterium-mediated transformation isefficient, it is the method of choice because of the facile and definednature of the gene locus transfer. The use of Agrobacterium-mediatedplant integrating vectors to introduce DNA into plant cells is wellknown in the art (Fraley et al., 1985; U.S. Pat. No. 5,563,055).

Transformation of plant protoplasts also can be achieved using methodsbased on calcium phosphate precipitation, polyethylene glycol treatment,electroporation, and combinations of these treatments (see, e.g.,Potrykus et al., 1985; Omirulleh et al., 1993; Fromm et al., 1986;Uchimiya et al., 1986; Marcotte et al., 1988). Transformation of plantsand expression of foreign genetic elements is exemplified in Choi et al.(1994), and Ellul et al. (2003).

A number of promoters have utility for plant gene expression for anygene of interest including but not limited to selectable markers,scoreable markers, genes for pest tolerance, disease resistance,nutritional enhancements and any other gene of agronomic interest.Examples of constitutive promoters useful for plant gene expressioninclude, but are not limited to, the cauliflower mosaic virus (CaMV)P-35S promoter, which confers constitutive, high-level expression inmost plant tissues (see, e.g., Odel et al., 1985), including in monocots(see, e.g., Dekeyser et al., 1990; Terada and Shimamoto, 1990); atandemly duplicated version of the CaMV 35S promoter, the enhanced 35Spromoter (P-e35S) the nopaline synthase promoter (An et al., 1988), theoctopine synthase promoter (Fromm et al., 1989); and the figwort mosaicvirus (P-FMV) promoter as described in U.S. Pat. No. 5,378,619 and anenhanced version of the FMV promoter (P-eFMV) where the promotersequence of P-FMV is duplicated in tandem, the cauliflower mosaic virus19S promoter, a sugarcane bacilliform virus promoter, a commelina yellowmottle virus promoter, and other plant DNA virus promoters known toexpress in plant cells.

A variety of plant gene promoters that are regulated in response toenvironmental, hormonal, chemical, and/or developmental signals can alsobe used for expression of an operably linked gene in plant cells,including promoters regulated by (1) heat (Callis et al., 1988), (2)light (e.g., pea rbcS-3A promoter, Kuhlemeier et al., 1989; maize rbcSpromoter, Schaffner and Sheen, 1991; or chlorophyll a/b-binding proteinpromoter, Simpson et al., 1985), (3) hormones, such as abscisic acid(Marcotte et al., 1989), (4) wounding (e.g., wunl, Siebertz et al.,1989); or (5) chemicals such as methyl jasmonate, salicylic acid, orSafener. It may also be advantageous to employ organ-specific promoters(e.g., Roshal et al., 1987; Schernthaner et al., 1988; Bustos et al.,1989).

Exemplary nucleic acids which may be introduced to plants of thisinvention include, for example, DNA sequences or genes from anotherspecies, or even genes or sequences which originate with or are presentin the same species, but are incorporated into recipient cells bygenetic engineering methods rather than classical reproduction orbreeding techniques. However, the term “exogenous” is also intended torefer to genes that are not normally present in the cell beingtransformed, or perhaps simply not present in the form, structure, etc.,as found in the transforming DNA segment or gene, or genes which arenormally present and that one desires to express in a manner thatdiffers from the natural expression pattern, e.g., to over-express.Thus, the term “exogenous” gene or DNA is intended to refer to any geneor DNA segment that is introduced into a recipient cell, regardless ofwhether a similar gene may already be present in such a cell. The typeof DNA included in the exogenous DNA can include DNA which is alreadypresent in the plant cell, DNA from another plant, DNA from a differentorganism, or a DNA generated externally, such as a DNA sequencecontaining an antisense message of a gene, or a DNA sequence encoding asynthetic or modified version of a gene.

Many hundreds if not thousands of different genes are known and couldpotentially be introduced into a broccoli plant according to theinvention. Non-limiting examples of particular genes and correspondingphenotypes one may choose to introduce into a broccoli plant include oneor more genes for insect tolerance, such as a Bacillus thuringiensis(B.t.) gene, pest tolerance such as genes for fungal disease control,herbicide tolerance such as genes conferring glyphosate tolerance, andgenes for quality improvements such as yield, nutritional enhancements,environmental or stress tolerances, or any desirable changes in plantphysiology, growth, development, morphology or plant product(s). Forexample, structural genes would include any gene that confers insecttolerance including but not limited to a Bacillus insect control proteingene as described in WO 99/31248, herein incorporated by reference inits entirety, U.S. Pat. No. 5,689,052, herein incorporated by referencein its entirety, U.S. Pat. Nos. 5,500,365 and 5,880,275, hereinincorporated by reference in their entirety. In another embodiment, thestructural gene can confer tolerance to the herbicide glyphosate asconferred by genes including, but not limited to Agrobacterium strainCP4 glyphosate resistant EPSPS gene (aroA:CP4) as described in U.S. Pat.No. 5,633,435, herein incorporated by reference in its entirety, orglyphosate oxidoreductase gene (GOX) as described in U.S. Pat. No.5,463,175, herein incorporated by reference in its entirety.

Alternatively, the DNA coding sequences can affect these phenotypes byencoding a non-translatable RNA molecule that causes the targetedinhibition of expression of an endogenous gene, for example viaantisense- or cosuppression-mediated mechanisms (see, for example, Birdet al., 1991). The RNA could also be a catalytic RNA molecule (i.e., aribozyme) engineered to cleave a desired endogenous mRNA product (seefor example, Gibson and Shillito, 1997). Thus, any gene which produces aprotein or mRNA which expresses a phenotype or morphology change ofinterest is useful for the practice of the present invention.

D. Definitions

In the description and tables herein, a number of terms are used. Inorder to provide a clear and consistent understanding of thespecification and claims, the following definitions are provided:

Allele: Any of one or more alternative forms of a gene locus, all ofwhich alleles relate to one trait or characteristic. In a diploid cellor organism, the two alleles of a given gene occupy corresponding locion a pair of homologous chromosomes.

Backcrossing: A process in which a breeder repeatedly crosses hybridprogeny, for example a first generation hybrid (F₁), back to one of theparents of the hybrid progeny. Backcrossing can be used to introduce oneor more single locus conversions from one genetic background intoanother.

Crossing: The mating of two parent plants.

Cross-pollination: Fertilization by the union of two gametes fromdifferent plants.

Diploid: A cell or organism having two sets of chromosomes.

Emasculate: The removal of plant male sex organs or the inactivation ofthe organs with a cytoplasmic or nuclear genetic factor or a chemicalagent conferring male sterility.

Enzymes: Molecules which can act as catalysts in biological reactions.

F₁ Hybrid: The first generation progeny of the cross of two nonisogenicplants.

Genotype: The genetic constitution of a cell or organism.

Haploid: A cell or organism having one set of the two sets ofchromosomes in a diploid.

Linkage: A phenomenon wherein alleles on the same chromosome tend tosegregate together more often than expected by chance if theirtransmission was independent.

Marker: A readily detectable phenotype, preferably inherited incodominant fashion (both alleles at a locus in a diploid heterozygoteare readily detectable), with no environmental variance component, i.e.,heritability of 1.

Phenotype: The detectable characteristics of a cell or organism, whichcharacteristics are the manifestation of gene expression.

Quantitative Trait Loci (QTL): Quantitative trait loci (QTL) refer togenetic loci that control to some degree numerically representabletraits that are usually continuously distributed.

Resistance: As used herein, the terms “resistance” and “tolerance” areused interchangeably to describe plants that show no symptoms to aspecified biotic pest, pathogen, abiotic influence or environmentalcondition. These terms are also used to describe plants showing somesymptoms but that are still able to produce marketable product with anacceptable yield. Some plants that are referred to as resistant ortolerant are only so in the sense that they may still produce a crop,even though the plants are stunted and the yield is reduced.

Regeneration: The development of a plant from tissue culture.

Royal Horticultural Society (RHS) color chart value: The RHS color chartis a standardized reference which allows accurate identification of anycolor. A color's designation on the chart describes its hue, brightnessand saturation. A color is precisely named by the RHS color chart byidentifying the group name, sheet number and letter, e.g., Yellow-OrangeGroup 19A or Red Group 41B.

Self-pollination: The transfer of pollen from the anther to the stigmaof the same plant.

Single Locus Converted (Conversion) Plant: Plants which are developed bya plant breeding technique called backcrossing, wherein essentially allof the desired morphological and physiological characteristics of abroccoli variety are recovered in addition to the characteristics of thesingle locus transferred into the variety via the backcrossing techniqueand/or by genetic transformation.

Substantially Equivalent: A characteristic that, when compared, does notshow a statistically significant difference (e.g., p=0.05) from themean.

Tissue Culture: A composition comprising isolated cells of the same or adifferent type or a collection of such cells organized into parts of aplant.

Transgene: A genetic locus comprising a sequence which has beenintroduced into the genome of a broccoli plant by transformation.

E. Deposit Information

A deposit was made of at least 2500 seeds of broccoli hybrid RX05991199. The deposit was made with the American Type Culture Collection(ATCC), 10801 University Boulevard, Manassas, Va. 20110-2209 USA. Thedeposit is assigned ATCC Accession No. PTA-13165. The date of depositwas Aug. 24, 2012. Access to the deposits will be available during thependency of the application to persons entitled thereto upon request.The deposits will be maintained in the ATCC Depository, which is apublic depository, for a period of 30 years, or 5 years after the mostrecent request, or for the enforceable life of the patent, whichever islonger, and will be replaced if nonviable during that period. Applicantdoes not waive any infringement of their rights granted under thispatent or any other form of variety protection, including the PlantVariety Protection Act (7 U.S.C. 2321 et seq.).

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity andunderstanding, it will be obvious that certain changes and modificationsmay be practiced within the scope of the invention, as limited only bythe scope of the appended claims.

All references cited herein are hereby expressly incorporated herein byreference.

REFERENCES

The following references, to the extent that they provide exemplaryprocedural or other details supplementary to those set forth herein, arespecifically incorporated herein by reference:

-   U.S. Pat. No. 5,378,619-   U.S. Pat. No. 5,463,175-   U.S. Pat. No. 5,500,365-   U.S. Pat. No. 5,563,055-   U.S. Pat. No. 5,633,435-   U.S. Pat. No. 5,689,052-   U.S. Pat. No. 5,880,275-   An et al., Plant Physiol., 88:547, 1988.-   Bird et al., Biotech. Gen. Engin. Rev., 9:207, 1991.-   Bustos et al., Plant Cell, 1:839, 1989.-   Callis et al., Plant Physiol., 88:965, 1988.-   Choi et al., Plant Cell Rep., 13: 344-348, 1994.-   Dekeyser et al., Plant Cell, 2:591, 1990.-   Ellul et al., Theor. Appl. Genet., 107:462-469, 2003.-   EP 534 858-   Fraley et al., Bio/Technology, 3:629-635, 1985.-   Fromm et al., Nature, 312:791-793, 1986.-   Fromm et al., Plant Cell, 1:977, 1989.-   Gibson and Shillito, Mol. Biotech., 7:125,1997-   Klee et al., Bio-Technology, 3(7):637-642, 1985.-   Kuhlemeier et al., Plant Cell, 1:471, 1989.-   Marcotte et al., Nature, 335:454, 1988.-   Marcotte et al., Plant Cell, 1:969, 1989.-   Odel et al., Nature, 313:810, 1985.-   Omirulleh et al., Plant Mol. Biol., 21(3):415-428, 1993.-   Potrykus et al., Mol. Gen. Genet., 199:183-188, 1985.-   Roshal et al., EMBO J., 6:1155, 1987.-   Schaffner and Sheen, Plant Cell, 3:997, 1991.-   Schernthaner et al., EMBO J., 7:1249, 1988.-   Siebertz et al., Plant Cell, 1:961, 1989.-   Simpson et al., EMBO J., 4:2723, 1985.-   Terada and Shimamoto, Mol. Gen. Genet., 220:389, 1990.-   Uchimiya et al., Mol. Gen. Genet., 204:204, 1986.-   Wang et al., Science, 280:1077-1082, 1998.-   Williams et al., Nucleic Acids Res., 1 8:6531-6535, 1990.-   WO 99/31248

What is claimed is:
 1. A seed of broccoli hybrid RX 05991199, a sampleof seed of said hybrid having been deposited under ATCC Accession NumberPTA-13165.
 2. A plant grown from the seed of claim
 1. 3. A plant part ofthe plant of claim
 2. 4. The plant part of claim 3, wherein said part isselected from the group consisting of a fruit, an ovule, pollen, a leaf,or a cell.
 5. A broccoli plant, or a part thereof, having all thephysiological and morphological characteristics of the broccoli plant ofclaim
 2. 6. A tissue culture of regenerable cells of broccoli hybrid RX05991199, a sample of seed of said hybrid having been deposited underATCC Accession Number PTA-13165.
 7. The tissue culture according toclaim 6, comprising cells or protoplasts from a plant part selected fromthe group consisting of embryos, meristems, cotyledons, pollen, leaves,anthers, roots, root tips, pistil, flower, seed, florets and stalks. 8.A broccoli plant regenerated from the tissue culture of claim 6, whereinthe regenerated plant expresses all of the physiological andmorphological characteristics of broccoli hybrid RX 05991199, a sampleof seed of said hybrid having been deposited under ATCC Accession NumberPTA-13165.
 9. A method of producing broccoli seed, comprising crossingthe plant of claim 2 with itself or a second broccoli plant.
 10. Amethod for producing a seed of a hybrid RX 05991199-derived broccoliplant comprising the steps of: (a) crossing a broccoli plant of hybridRX 05991199, a sample of seed of said hybrid having been deposited underATCC Accession Number PTA-13165, with a second broccoli plant; and (b)allowing seed of a RX 05991199-derived broccoli plant to form.
 11. Themethod of claim 10, further comprising the steps of: (c) selfing theplant grown from said RX 05991199-derived broccoli seed or crossing itto a second broccoli plant to yield additional RX 05991199-derivedbroccoli seed; (d) growing said additional RX 05991199-derived broccoliseed of step (c) to yield additional RX 05991199-derived broccoliplants; and (e) repeating the steps of (c) and (d) to generate furtherRX 05991199-derived broccoli plants.
 12. A method of vegetativelypropagating a plant of broccoli hybrid RX 05991199 comprising the stepsof: (a) collecting tissue capable of being propagated from a plant ofbroccoli hybrid RX 05991199, a sample of seed of said hybrid having beendeposited under ATCC Accession Number PTA-13165; (b) cultivating saidtissue to obtain proliferated shoots; and (c) rooting said proliferatedshoots to obtain rooted plantlets.
 13. The method of claim 12, furthercomprising growing plants from said rooted plantlets.
 14. A method ofproducing a plant of broccoli hybrid variety RX 05991199 comprising anadded desired trait, the method comprising introducing a transgeneconferring the desired trait into broccoli hybrid RX 05991199, whereby aplant of broccoli hybrid RX 05991199 comprising an added desired traitis produced, a sample of seed of said variety having been depositedunder ATCC Accession Number PTA-13165.
 15. A plant of broccoli hybridvariety RX 05991199 comprising a transgene conferring a desired trait, asample of seed of said variety has been deposited under ATCC AccessionNumber PTA-13165.
 16. A hybrid broccoli plant that exhibits all of thetraits of broccoli hybrid RX 05991199, a sample of seed of said hybridhaving been deposited under ATCC Accession Number PTA-13165.
 17. A seedthat produces the plant of claim
 15. 18. A seed that produces the plantof claim
 16. 19. A method of producing a broccoli plant part comprising:(a) obtaining the plant of claim 1, wherein the plant has beencultivated to maturity; and (b) collecting at least a first plant partfrom the plant.
 20. The method of claim 19, wherein the plant partcomprises a floret.